At a fundamental level, food is viewed as a source of nutrition to meet daily requirements at a minimum in order to survive but with an ever greater focus on the desire to thrive. In the latter instance, there is an ever growing interest in the functionality of food. Functional foods have been defined as any modified food or food ingredient that may provide a health benefit beyond the traditional nutrients it contains. The term nutraceutical is defined as “any substance that may be considered a food or part of a food and provides health benefits, including the prevention and treatment of disease”. Further to it, not only productivity of crop but amino acid content, mineral content is of crucial importance ultimately for human health who consumes it.
Protein energy malnutrition is the most lethal form of malnutrition and affects every fourth child worldwide. The Food and Agriculture Organization estimates that 850 million people worldwide suffer from under nutrition, to which insufficient protein in the diet is a significant contributing factor. Most plants have a poor balance of essential amino acids relative to the needs of animals and humans. The cereals such as maize, wheat [Triticum aestivum], rice, etc. tend to be low in Lysine, whereas legumes such as soybean and pea [Pisum sativum] are often low in the sulfur-rich amino acids Methionine and Cysteine. Micronutrient malnutrition, the so-called hidden hunger, affects more than half of the world's population, especially women and preschool children in developing countries (United Nations System Standing Committee on Nutrition (2004) United Nations System Standing Committee on Nutrition 5th Report on the World Nutrition Situation: Nutrition for Improved Development Outcomes. Higher level of micronutrients in plants is one way of increasing the possibility of their higher intake while consumption.
Agricultural innovation has always involved new, science-based products and processes that have contributed reliable methods for increasing productivity and sustainability. Biotechnology has introduced a new dimension to such innovation, offering efficient and cost-effective means to produce a diverse array of novel, value-added products and tools. Plant foods can serve as dietary sources of all essential amino acids and minerals required by humans. Amino acids and mineral concentrations are low in some plants, especially many staple food crops; thus, efforts are underway to increase the amino acids and minerals content of these foods as a means to ensure adequate attainment of dietary minerals in all individuals. While these efforts have included classical breeding approaches in the past, it is clear that future progress can be made by utilizing the tools of biotechnology to effect directed changes in plant mineral status. Different approaches have been developed to overcome the problem of low level of minerals, amino acids and increase in productivity of plants. Value-added output traits, such as improved nutrition and food functionality and plants as production factories for therapeutics and industrial products from a consumer perspective, the focus on value added traits, especially improved nutrition, is of greatest interest (Martina Newell-McGloughlin (2008) Nutritionally Improved Agricultural Crops. Plant Physiology, 147, 939-953). Trichoderma spp. is an asexually reproducing, free-living fungi that is common in soil and root ecosystems. It is one of the most exploited fungal biocontrol agents in the field of agriculture for the management of crop diseases caused by a wide range of fungal phytopathogens. Trichoderma species have been investigated as biological control agents for over 75 years, but it is only recently that isolates have become commercially available. These organisms have been favored because they are able to control a wide variety of phytopathogenic fungi that are of great importance to agriculture. Trichoderma spp. can control a wide variety of pathogens and appear in more products than any other microbe including Anti-Fungus; Binab T; Supresivit; T-22G and T-22HB; Trichopel, Trichoject, Trichodowels, and Trichoseal; TY. Products containing Trichoderma spp. control species of Amillaria, Botrytis, Chondrostrenum, Colletotrichum, Fulvia, Fusarium, Monilia, Nectria, Phytophthora, Plasmopara, Psendoperonospora, Pythium, Rhizoctonia, Rhizopus, Sclerotinia sclerotiorum, Sclerotium rolfsii, Verticillium, and wood rot fungi.
Though the available strains of Trichoderma obtained through selection or mutation, possess one or more desirable traits, none of them possess all the attributes to realize the full potential of the beneficial fungus. One method of combining characteristics from different fungi is protoplast fusion. Protoplast fusion allows the transfer of complex traits without having to know the genes involved, and for genetic recombination between organisms that cannot undergo sexual recombination. Therefore, the protoplast fusion technology has stimulated interest in the manipulation of Trichoderma as enzyme producers and bio-control agents against diverse plant pathogens (Hanson E. L. and C. R. Howell. (2002). Biocontrol efficacy and other characteristics of protoplast fusants between Trichoderma koningii and T. virens. Mycol. Res. 106:321-328).
While work on Trichoderma has been conducted in the past, there is no clear indication heretofore that any detailed study has been conducted to demonstrate the composition of Trichoderma showing increase in amino acid, trace elements, increase in chlorophyll content and yield simultaneously in plants, using this technique. Thus, there exists a problem in prior art where composition of biological agent needs to be developed for effecting increase in amino acid, trace elements, increase in chlorophyll content and yield simultaneously in plants. For the purpose, improved strains of Trichoderma are needed to be developed. Protoplast fusion of parent strains offers one option for the development of novel strains. Therefore, the present work is aimed to isolate protoplasts from parent Trichoderma strains followed by self-fusion of protoplasts for investigating the possible enhancement of amino acids, trace elements, an increase in chlorophyll content, and yield of plants. In the present invention, solution to the problems of prior art have been provided, by employing biotechnology as a tool to enhance the amino acids, trace elements, chlorophyll content, and yield of plants.